目的 探討人參皂苷F₂（GF₂）對α-萘異硫氰酸酯（ANIT）誘導(dǎo)的膽汁淤積肝損傷（CLI）小鼠的作用及機(jī)制。方法 將70只雄性昆明小鼠隨機(jī)分為7組（n＝10）：對照組、單給GF₂（100 mg∙kg^-1）組、模型組、熊去氧膽酸（UDCA，40 mg∙kg^-1）組和GF₂低、中、高劑量（25、50、100 mg∙kg^-1）組。小鼠連續(xù)ig給藥7 d，于第5天ig給予ANIT（100 mg∙kg^-1）建立膽汁淤積模型。自動生化儀測量血清丙氨酸氨基轉(zhuǎn)移酶（ALT）、天冬氨酸氨基轉(zhuǎn)移酶（AST）、堿性磷酸酶（ALP）、總膽汁酸（TBA）、總膽紅素（TBIL）、直接膽紅素（DBIL）、丙二醛（MDA）、超氧化物歧化酶（SOD）、谷胱甘肽過氧化物酶（GSH-Px）水平，肝組織勻漿上清MDA、SOD、GSH-Px和過氧化氫酶（CAT）水平；ELISA法檢測肝組織勻漿上清液中炎癥因子腫瘤壞死因子-α（TNF-α）、白細(xì)胞介素（IL）-6、IL-1β和脂多糖（LPS）水平；HE染色和Masson染色進(jìn)行肝組織病理學(xué)分析；TUNEL染色觀察肝細(xì)胞凋亡；免疫組化法觀察免疫細(xì)胞標(biāo)志物[中性粒細(xì)胞標(biāo)志物CD11b和Ly6g、巨噬細(xì)胞標(biāo)志物F4/80和T細(xì)胞標(biāo)志物CD3]、肝纖維化標(biāo)志物[α-平滑肌肌動蛋白（α-SMA）、I型膠原（Collagen I）]、Toll樣受體4（TLR4）/髓分化因子88（MyD88）/核因子κB（NF-κB）相關(guān)蛋白、轉(zhuǎn)化生長因子-β1（TGF-β1）/Smad相關(guān)蛋白、Bcl-2相關(guān)X蛋白（Bax）蛋白表達(dá)；Western blotting檢測TLR4/Myd88/NF-κB、Nrf2/HO-1/NQO1和TGF-β1/Smad、B淋巴細(xì)胞瘤-2（Bcl-2）/Bax通路蛋白表達(dá)。結(jié)果 與模型組比較，GF₂組AST、ALT、ALP、TBA、TBIL和DBIL水平顯著降低（P＜0.05、0.01、0.001），GF₂明顯改善模型組肝細(xì)胞腫脹、空泡化、肝內(nèi)炎癥細(xì)胞浸潤和壞死，膽管增生和擴(kuò)張；GF₂顯著降低ANIT誘導(dǎo)的炎癥因子TNF-α、IL-6、IL-1β、LPS水平（P＜0.05、0.01、0.001），明顯降低CD11b、Ly6g、F4/80、CD3表達(dá)，顯著降低TLR4、Myd88、NF-κB p65和pIκBα的蛋白表達(dá)（P＜0.05、0.01）；GF₂組MDA水平降低，SOD、CAT活性和GSH水平恢復(fù)，差異顯著（P＜0.05、0.01、0.001），且上調(diào)Nrf2及其靶基因HO-1和NQO1蛋白的表達(dá)（P＜0.05、0.001）；GF₂組肝纖維化明顯減輕，α-SMA和Collagen I表達(dá)明顯下降，且TGF-β1、Smad2、Smad3蛋白表達(dá)顯著降低（P＜0.05、0.01、0.001）；GF₂減弱CLI小鼠的肝細(xì)胞凋亡程度，顯著降低促凋亡蛋白Bax的表達(dá)、顯著增加抗凋亡蛋白Bcl-2的表達(dá)（P＜0.05、0.01）。結(jié)論 CF₂可抑制TLR4/Myd88/NF-κB通路減輕炎癥反應(yīng)、激活Nrf2通路減輕小鼠氧化損傷、抑制TGF-β1/Smad通路減輕肝纖維化，減輕ANIT誘導(dǎo)的細(xì)胞凋亡，進(jìn)而減輕膽汁淤積小鼠肝損傷。

Objective To investigate the effect of ginsenoside F₂(GF₂) on α-naphthalene isothiocyanate (ANIT) -induced cholestatic liver injury (CLI) in mice and its mechanism. Methods 70 male Kunming mice were randomly divided into seven groups (n= 10): control group, single GF₂ (100 mg∙kg^-1) group, model group, UDCA (40 mg∙kg^-1) group, and low, medium, and high dose GF₂ (25, 50, 100 mg∙kg^-1) groups. The mice were administered orally with the drugs for seven consecutive days, and ANIT (100 mg∙kg^-1) was ig administered on the 5th day to establish a cholestasis model. The automatic biochemistry analyzer was used to measure serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), total bile acid (TBA), total bilirubin (TBIL), direct bilirubin (DBIL), malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) levels, and the levels of MDA, SOD, GSH-Px, and catalase in the liver tissue homogenate supernatant; the enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of inflammatory factors (tumor necrosis factor-α (TNF-α), interleukin (IL)-6, IL-1β, and LPS) in the liver tissue homogenate supernatant; hematoxylin and Masson staining were used for histopathological analysis of the liver tissue; TUNEL staining was used to observe hepatocyte apoptosis; immunohistochemistry was used to detect the expression of immune cell markers (neutrophil markers CD11b and Ly6g, macrophage markers F4/80 and T cell markers CD3), liver fibrosis markers (α-smooth muscle actin (α-SMA), type I collagen (Collagen I)), Toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear factor kappa B (NF-κB) related protein, transforming growth factor-β1 (TGF-β1)/SMAD-associated protein, Bcl-2 associated X protein (Bax) protein expression. Western blotting was used to detect the expression of TLR4/Myd88/NF-κB, Nrf2/HO-1/NQO1, TGF-β1/Smad, B-lymphoblastoma-2 (Bcl-2)/Bax pathway proteins. Results Compared with the model group, the levels of AST, ALT, ALP, TBA, TBIL and DBIL were significantly lower in the GF₂ group (P < 0.05, 0.01, 0.001), and GF₂ significantly improved the liver cell swelling, vacuolization, intrahepatic inflammatory cell infiltration and necrosis, bile duct hyperplasia and dilation in the model group; GF₂ significantly lowered the levels of inflammatory factors TNF-α, IL-6, IL-1β and LPS induced by ANIT (P < 0.05, 0.01, 0.001), and significantly lowered the expression of CD11b, Ly6g, F4/80, CD3, and significantly downregulated the protein expression of TLR4, Myd88, NF-κB p65 and pIκBα (P < 0.05, 0.01); the level of MDA was lowered in the GF₂ group, and the activities of SOD and CAT and the level of GSH were restored, with significant differences (P < 0.05, 0.01, 0.001), and the expression of Nrf2 and its target genes HO-1 and NQO1 was significantly upregulated (P < 0.05, 0.001); the degree of liver fibrosis was significantly alleviated in the GF₂ group, and the expression of α-SMA and Collagen I was significantly downregulated, and the expression of TGF-β1, Smad2 and Smad3 was significantly downregulated (P < 0.05, 0.01, 0.001); GF₂ weakened the degree of liver cell apoptosis in CLI mice, significantly decreased the expression of pro-apoptotic protein Bax and significantly increased the expression of anti-apoptotic protein Bcl-2 (P < 0.05, 0.01). Conclusion CF₂ can inhibit the TLR4/Myd88/NF-κB pathway to alleviate inflammatory responses, activate the Nrf2 pathway to alleviate oxidative damage in mice, inhibit the TGF-β1/Smad pathway to alleviate liver fibrosis, alleviate cell apoptosis induced by ANIT, and thereby alleviate liver damage in cholestasis mice.

2024年度河南省中醫(yī)藥科學(xué)研究專項重點項目（2024ZY1030）